The effect of various concentration of methrotrexate on the interleukin-17 from peripheral blood mononuclear cells in vitro
10.3760/cma.j.issn.1007-7480.2010.08.006
- VernacularTitle:不同浓度甲氨蝶呤对外周血单个核细胞表达和分泌白细胞介素-17的影响
- Author:
Yanshan LI
;
Lindi JIANG
;
Si ZHANG
;
Lianhua YIN
;
Lili MA
;
Huiyong CHEN
;
Zhen WANG
- Publication Type:Journal Article
- Keywords:
Interleukin-17;
Methotrexate;
Arthritis,rheumatoid;
Peripheral blood mononuclear cell
- From:
Chinese Journal of Rheumatology
2010;14(8):535-537
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of different concentrations of methotrexate (MTX) on IL-17 from peripheral blood mononuclear cells(PBMCs) and To clarify the active mechanisms of MTX on RA. Methods PBMCs were isolated from heparinized blood of healthy donors or patients with RA using Ficoll-Hypaque density gradient centrifugation. The cells were pretreated with various concentrations of MTX and then stimulated by anti-human CD3/anti-human CD28 at 37℃5%CO2. The IL-17 mRAN level was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The supernatants were harvested and the protein level of IL-17 was tested by ELISA kit. The percentage of CD4+IL-17+cells in PBMCs was detected by flow cytometry. Results For the four different concentrations of MTX groups (0.1,1.0, 5, 25μg/ml), the IL-17 mRNA/GAPDH ratio(0.58±0.09,0.48±0.11, 0.50±0.09, 0.51±0.14) were lower than those of the non-drug group(0.76±0.08). Paired-t test or independent-samplet test showed significant difference between the MTX treatment group and the non-drug group(P＜0.01). The level of IL-17 of the four MTX groups was(121±54)pg/ml and(104±45)pg/ml and(90±36)pg/ml and(115±41)pg/ml, which was lower than the non-drug group(370±187)pg/ml(P＜0.01). The average C D4+IL-17+cell ratio was reduced, but had no statistically signficant differences(P＞0.05). Conclusion MTX can decrease Th17 cells differentiation and suppress IL-17 production of PBMCs, but no association can be found between its effect on the expression of IL-17 and the concentration of MTX.
